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Analytical considerations for reducing the matrix effect for the sphingolipidome quantification in whole blood
Dezhen Wang1, Peining Xu1, Clementina Mesaros1
1Department of Systems Pharmacology & Translational Therapeutics, University of Pennsylvania, Philadelphia, PA 19104, USA.
Bioanalysis
|June 10, 2021
Summary
Whole blood (WB) analysis offers a more comprehensive view of the sphingolipidome than plasma. This method quantizes over 150 sphingolipids, revealing lower variability and higher concentrations in WB for biomarker discovery.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biomarker Discovery
Background:
- Plasma and serum are common biofluids for metabolomics and lipidomics.
- High-concentration analytes in blood cells are missed in plasma/serum assays.
- Sample isolation can introduce variability into lipidomics data.
Purpose of the Study:
- To develop a comprehensive method for whole blood (WB) sphingolipidome quantification.
- To compare sphingolipid levels and variability between WB and plasma.
- To assess WB as a superior alternative to plasma for lipidomics and biomarker discovery.
Main Methods:
- A single-phase extraction technique was employed.
- Liquid chromatography-high-resolution mass spectrometry (LC-HRMS) was utilized for quantification.
- Paired whole blood and plasma samples were analyzed.
Main Results:
- Over 150 sphingolipids were successfully quantified in whole blood.
- Most sphingolipids were found at higher concentrations in WB compared to plasma.
- Individual variations in sphingolipid levels were lower in WB.
Conclusions:
- Whole blood is a more informative biofluid than plasma for sphingolipidome analysis.
- WB analysis reduces variability and captures a broader range of analytes.
- This method can enhance sphingolipid biomarker discovery.
